Localization of platelet-derived growth factor (PDGF) in CHO cells transfected with PDGF A- or B-chain cDNA: retention of PDGF-BB in the endoplasmic reticulum and Golgi complex.
Platelet-derived growth factor (PDGF) is a powerful mitogen for connective tissue cells. It is made up of two polypeptide chains (A and B) and exists in three dimeric forms (AA, AB, and BB). Transfection experiments have indicated that PDGF-AA and -AB are secreted as 30 x 10(3) Mr products, whereas PDGF-BB is processed into a 24 x 10(3) Mr product and remains associated with the cells. Here, CHO cells were transfected with PDGF B- or A-chain cDNA and the intracellular distributions of the respective gene products were compared by indirect immunofluorescence and immunoelectron microscopy, using primary antibodies specific for PDGF B- and A-chain homodimers. PDGF-BB was most conspicuous in stacked Golgi cisternae. It was also found in the endoplasmic reticulum and in lysosomes. Upon treatment of the cells with the microtubule-disruptive drug nocodazole, the Golgi complex was broken up and its stacks of cisternae were dispersed throughout the cytoplasm together with clusters of lysosomes. After this structural disorganization, the concentration of PDGF-BB to the Golgi stacks was even more prominent than before. Weak reactivity for PDGF-AA was detected in the endoplasmic reticulum and groups of vacuoles, both in control and nocodazole-treated cells, whereas Golgi stacks and lysosomes only seldom were positive. The observations suggest that PDGF-BB is processed and retained within the endoplasmic reticulum and Golgi complex. Eventually, it may also be transferred to lysosomes for degradation. In contrast, PDGF-AA is likely to follow a pathway for bulk flow, including rapid passage through the endoplasmic reticulum and Golgi complex, package in secretory vacuoles, and extracellular release by exocytosis.
Thyberg
J
J
Department of Medical Cell Biology, Karolinska Institutet, Stockholm, Sweden.
Ostman
A
A
Bäckström
G
G
Westermark
B
B
Heldin
C H
CH
D000818:Animals; D002460:Cell Line; D006224:Cricetinae; D003412:Cricetulus; D004721:Endoplasmic Reticulum / Q000378:metabolism*; D005260:Female; D005455:Fluorescent Antibody Technique; D006056:Golgi Apparatus / Q000187:drug effects / Q000378:metabolism*; D006801:Humans; D007124:Immunoenzyme Techniques; D008247:Lysosomes / Q000187:drug effects / Q000378:metabolism; D016253:Microscopy, Immunoelectron; D015739:Nocodazole / Q000494:pharmacology; D010053:Ovary / Q000166:cytology; D010982:Platelet-Derived Growth Factor / Q000235:genetics / Q000378:metabolism*; D011994:Recombinant Proteins / Q000378:metabolism; D014162:Transfection
D010982:Platelet-Derived Growth Factor; D011994:Recombinant Proteins; D015739:Nocodazole